1. Technical Field
The present invention relates to molecular biology and microbiology and, more particularly, to pregnancy tests for mammals and sex determination in animals. Still more particularly, this invention relates to pregnancy tests for equine mammals which are based on saliva or other bodily fluids.
2. Background Information
The prior art discloses a number of ways to test for pregnancy in female mammals. U.S. Pat. No. 3,968,011 to Manantou, et al., for example, discloses a method for clorimetrically assaying the quantity of N-acetyl-.beta.-glucosaminidase in a female biological medium, such as saliva, which quantity is indicia of fertility or pregnancy. The implement is an absorbent material, such as paper strip, impregnated with a phenolic derivative of N-acetyl-.beta.-d-glucosamine that reacts in the presence of the glucosaminidase at an acide pH to form a phenol that has a distinct color at an alkaline pH, and a buffer that maintains said acid pH. The method may be carried out by wetting the implement with the medium, allowing the phenol to form, raising the pH to alkalinity by wetting the implement with an appropriate buffer solution, and comparing the color of the implement with a color standard.
U.S. Pat. No. 4,003,988 to Hoff, et al. discloses a direct agglutination reagent for pregnancy testing which comprises the use of suspensions of polystryrene latex particles sensitized with a globulin fraction of anti-serum to human chorionic gonadotropin (HCG). When mixed with urine or blood serum samples containing HCG, this reagent agglutinates indicating a positive test for pregnancy.
U.S. Pat. No. 4,123,504 to Banik, et al. discloses a method and device for detecting pregnancy. This test involves concentration by ultrafiltration of a sample of urine or serum from a subject; followed by determining the presence of human chorionic gonadotropin or of its .beta.-subunit in the concentrated sample.
U.S. Pat. No. 4,348,207 to Cappel discloses a method for testing for pregnancy in humans in which a sample of a patient's first morning urine is added to a test tube containing a known lyophilized reagent. The tube is capped, shaken to mix the contents, and placed upright for one to two hours. The tube is then inverted and compared with positive and negative standard vials to show either agglutination of particles, such as red blood cells contained therein, in which case the subject is not pregnant, or a failure to agglutinate, in which event the patient is pregnant. The test tube is of sufficient dimension to support capillary action and is formed from, or has its interior surface coated with, a material which is non-wettable to the liquid contained therein.
U.S. Pat. No. 4,716,123 to Wood discloses a home pregnancy test in which a specifically binding biomaterial is attached to a macroextensive surface of a plastic strip or the like. A biological substance which is a specific binding partner to a binding site of the specifically binding biomaterial is attached to each of a plurality of synthetic particles. The particles are of a preselected size, refractive index, or the like to enhance their visibility in accordance with the Mie scattering phenomenon. Testing is by either contacting the particles with the strips to obtain adherence of the particles to the strips, or by exposing strips having the particles already adhering to them to a solution containing either the specifically binding biomaterial or the biological substance, whereby the particles adherence to the strip is eliminated.
U.S. Pat. No. 4,720,455 to Babu, et al. discloses a progesterone concentration level test for mammalian body fluids particularly adapted for milk whereby estrus and pregnancy can be determined. The test can be carried out with a kit of several reagents, test tubes and a dip-stick carrying an anti-progesterone monoclonal antibody.
U.S. Pat. No. 5,837,197 to Porrazzo, et al. discloses a fertility analysis and reproductive health system that is applicable to both female and male mammals. The invention disclosed in this patent is a portable, handheld, integrated unit which can be manufactured out of plastic. The unit can be disposable for hygienic purposes, or cleaned or sterilized for repeated use as desired. Aspects of the invention include the following: an immediate testing methodology employing any and all female fluids or secretions called positive fertility testing; the creation of a plastic, completely integrated, portable, self-contained and self-focusing examination system which relies on a visual reference system making it language independent; a test area section with replaceable slides where different, specific wavelengths of light are employed; the embodiment of a compound test areas so that multiple positive fertility tests may be conducted simultaneously; implementing the ability to immediately perform two or more positive fertility tests simultaneously using different female fluids or secretions; providing a novel batter powered microprocessor system to automatically perform the positive fertility testing; providing an accurate indicator of positive ovulation whereby the woman may pinpoint times of greatest fertility, thereby knowing the optimum time period for achieving pregnancy.
The prior art also discloses possibly relevant art concerning methods of detecting ovulation in female mammals. U.S. Pat. No. 4,385,125 to Preti, et al., for example, discloses that the level of certain alcohols is dramatically increased during the ovulation cycle of mammals. This patent thus utilizes this change to precisely ascertain the time of ovulation by monitoring the concentration of alcohols noting that a spike in the concentration indicates a time of ovulation. While this device and process of this patent determines the alcohol level by measuring such alcohol levels in saliva, it does not utilize reporter enzymes to measure the level of certain hormones in saliva to determine pregnancy, but only to measure alcohol to determine ovulation.
U.S. Pat. No. 5,460,976 to O'Connor discloses a method of predicting ovulation and a test kit is described which allows one to accurately predict the time of ovulation in an animal in advance thus permitting the highest rate of pregnancy to be achieved and at the same time minimizing embryonic death.
U.S. Pat. No. 5,721,142 to Klemm, et al. also relates to a method of monitoring the mammalian reproductive cycles. In the method disclosed in this patent the quantity of one or more low molecular weight compounds such as acetal/dehyde are monitored.
U.S. Pat. No. 5,837,197 to Porrazzo, et al. discloses a positive fertility testing and reproductive health system. Inasmuch as this patent relates generally to the measurement of reproductive health and fertility status, it discloses a number of the general features of the invention including testing for enzymes in saliva to determine a male or female's reproductive state and health.
U.S. Pat. No. 5,914,271 to Law, et al., discloses a fertility test and more particularly, relates to the testing of magnesium and calcium concentrations in saliva. More specifically, within the three to five days immediately preceding ovulation, the calcium and magnesium concentrations in the saliva drop. As a result, concentration monitoring can be done by any conventional means for quantitatively analyzing the calcium and magnesium to determine the fertility state of the user.
A need still exists, however, for improved ways of testing for pregnancy in mammals.
Many species of avians as well as other animals also do not display phenotypic differences between the sexes, especially in immature individuals. With respect to avians, such sex identification may be difficult in some commercial species such as parrots and ratites, and for certain endangered species such as the California Condor and Whooping Crane as well as non-endangered species, such as the Canada Goose.
Gender identification can be performed in sexually monomorphic birds and other animals through surgical examination. Alternatively, sex identification can sometimes be made through karyotype analysis.
Another alternative method of sex identification in birds is disclosed in U.S. Pat. No. 5,679,514 to Baker. This method comprises the steps of obtaining a DNA sequence which includes the W specific chromosome from the bird. Then, there is the step of identifying the nucleotides of the DNA sequence of the W specific chromosome. This prior art method also pertains to a method of determining the sex of a bird, and a method of determining genetic relatedness between two birds.
The above described methods of sex identification may, however, be relatively expensive and time consuming. A need, therefore, exists for an improved method of sex identification for monomorphic avians and other animals.